Detection of Glycosylation Sites (N-glycan, O-glycan) in Recombinant Protein Vaccines

Recombinant protein vaccines are a new type of vaccine product produced by genetic engineering technology in organisms or cell systems. During the production of these vaccines, glycosylation reactions may occur, that is, sugar chains are added to the N-sites (such as aspartic acid) and O-sites (such as serine or threonine) of proteins. Glycosylation is an important post-translational modification of proteins, affecting the stability, solubility, and biological activity of proteins. Therefore, for the quality control and efficacy evaluation of recombinant protein vaccines, it is essential to detect and analyze glycosylation sites.

 

There are many methods to detect glycosylation sites. The most common is mass spectrometry analysis, which can accurately determine the composition and linkage mode of sugar chains. This usually requires enzymatic digestion of proteins to produce peptides with glycosylation sites, and then these peptides are identified and quantified by mass spectrometry analysis. In addition, high-performance liquid chromatography is also an important tool. It can be used to separate and quantify sugar chains. By using specific fluorescent tags, sugar chains can be quantitatively analyzed. There are also more advanced technologies, such as nuclear magnetic resonance and X-ray crystallography, which can be used to study the detailed structure of glycosylation. Through these methods, scientists can deeply understand the glycosylation situation of vaccines, and then optimize the production process to improve the safety and effectiveness of vaccines.

 

In the analysis method of glycosylation sites of recombinant protein vaccines, mass spectrometry detection of glycosylation sites has extremely high sensitivity and can measure samples at femtomolar concentration levels. At the same time, mass spectrometry can be used to analyze complete and fragmented glycoprotein samples or peptides, and it can also detect and analyze unknown glycopeptides and glycoprotein compounds.

 

MtoZ Biolabs have two mass spectrometry analysis technologies, MALDI TOF MS and nano LC-ESI-MS/MS, to provide you with efficient and accurate identification services for glycosylation of recombinant protein vaccines. After receiving the sample, we first enzymatically digest the sample to release the Fc domain and Fab of the antibody, then separate the glycosylated fragments, and confirm the glycosylation sites through MALDI TOF MS or nano LC-ESI-MS/MS.